Cranial Motor Research Articles

Organization of acetylcholine containing structures in the cranial motor nuclei of the rhombencephalon of the pig

We explored the immunoreactivity of choline acetyltransferase (ChAT) in the cranial nerve motor nuclei of the porcine rhombencephalon to reveal the cholinergic nature of these regions. In our experiments we used an immunohistochemical method for the visualization of all acetylcholine-containing structures. All studied motor nuclei contained ChAT-positive cell bodies and fibres, but the intensity of staining differed between the nuclei. Furthermore, characteristic ChAT-immunoreactive bouton-like structures, which are known to be synaptic terminals of the cholinergic system, were observed in the borders of all studied regions. The localization of ChAT-positive “boutons” in the neuropil of the examined nuclei and their proximity to stained perikarya allowed us to differentiate two groups of motor nuclei in the rhombencephalon of the pig: (a) Nuclei containing ChAT-positive bouton-like structures dispersed in the neuropil, often establishing contacts with the stained cell bodies − motor trigeminal, abducent, facial, ambiguous and hypoglossal nuclei. (b) Nuclei in which characteristic boutons were dispersed among the ChAT-positive cells, but were devoid of any contact with perikarya − dorsal motor nucleus of the vagus nerve. These results provide new data on the porcine central nervous system and could be useful in further experiments on amyotrophic lateral sclerosis (ALS) − the disease that results in the progressive degeneration of motoneurons in the brain and spinal cord.

Palinacousis in temporal lobe intracerebral hemorrhage

Palinacousis, an auditory illusion in which perceived aural sensations persist or recur after the initial acoustic stimulus has ceased,1 may be caused by structural lesions involving the temporal lobe. However, the reported lesions were usually extensive, and detailed imaging tests were rarely performed.2-4 We report a patient with transient palinacousis following discrete temporal lobe intracerebral hemorrhage (ICH). A 67-year-old hypertensive woman suddenly developed language disturbances. On examination the next day, she was alert, and cranial nerves and motor and sensory functions were intact. Her speech was fluent and mixed with frequent paraphasic errors. She showed difficulty in understanding and naming. The Korean version of the Western Aphasia Battery confirmed that she had severe Wernicke-type aphasia (Aphasia Quotient = 27.2/100). Brain MRI showed an acute ICH in the left middle temporal gyrus (figure). Her language disturbances rapidly improved. Three days after the symptom onset, auditory comprehension (0.5/10 to 7.5/10) and repetition (3.1/10 to 6.7/10) considerably improved, but there remained significant naming deficits (0/10 to …

Does Electrode Placement Influence Quality of Intraoperative Monitoring of Cranial Motor Nerves?

Background: Continuous EMG monitoring is a standard method for intraoperative monitoring of motor cranial nerves. This method uses needle electrodes which are inserted into muscles innervated by the monitored nerves according to various setups. The aim of this study was to compare sensitivity and noise levels of different electrode setups using the example of facial nerve monitoring.

Specific and artifactual labeling in the rat spinal cord and medulla after injection of monosynaptic retrograde tracers into the diaphragm

The use of fluorescent dyes has been a major improvement for paths tracing studies. However, these tracers present different properties and have to be chosen carefully. The present study compares the ability of different tracers to specifically label phrenic motoneurons (PMNs) innervating the rat diaphragm. The administration of fluorogold (FG) from the transected phrenic nerve specifically labeled PMNs in the ipsilateral spinal cord. However, when FG was injected into one hemidiaphragm, in addition with ipsilateral PMNs, a less intense artifactual labeling was observed in the spinal cord (mainly in contralateral PMNs) and in the medulla oblongata (mainly in the area postrema and cranial motor nuclei). Similar results were observed using horseradish peroxidase, while no labeling was observed after injection of nuclear yellow or diamidino yellow into the diaphragm. By contrast, the dextran amine fluororuby (FR) and the carbocyanine DiAsp selectively and exclusively labeled ipsilateral PMNs 2 or 3 weeks after injection into the diaphragm, respectively. The lipophilic properties of DiAsp and the high molecular weight of FR may prevent their diffusion to adjacent tissues and into the blood stream which seems to account for the artifactual labeling observed with the other tracers. The higher homogeneity and quality of the labeling observed with FR compared to DiAsp make it the most appropriate tracer for the specific monosynaptic fluorescent labeling of PMNs after injection into the diaphragm.

Distribution and cellular localization of imidazoleacetic acid-ribotide, an endogenous ligand at imidazol(in)e and adrenergic receptors, in rat brain

Imidazoleacetic acid-ribotide (IAA-RP) is a putative neurotransmitter/modulator recently discovered in mammalian brain. The present study examines the distribution of IAA-RP in the rat CNS using a highly specific antiserum raised in rabbit against IAA-RP with immunostaining of aldehyde-fixed rat CNS. IAA-RP-immunoreactive neurons were present throughout the neuraxis; neuroglia were not labeled. In each region, only a subset of the neuronal pool was immunostained. In the forebrain, ribotide-immunolabeled neurons were common in neocortex, in hippocampal formation, and in subcortical structures including basal ganglia, thalamus and hypothalamus. Labeling was prominent in limbic areas including olfactory bulb, basal forebrain, pyriform cortex and amygdala. In the mid- and hindbrain, immunolabeled neurons were concentrated in specific nuclei and, in some areas, in specific subregions of those nuclei. Structures of the motor system, including cranial nerve motor nuclei, precerebellar nuclei, the substantia nigra, and the red nucleus were clearly labeled. Staining was intense in cells and/or puncta in the rostral and caudal ventrolateral medullary reticular formation, nucleus tractus solitarius and the caudal vestibular nuclear complex. Within neurons, the ribotide was found predominantly in somata and dendrites; some myelinated axons and occasional synaptic terminals were also immunostained. These data indicate that IAA-RP contributes to the neurochemical phenotype of many neuronal populations and further supports our suggestion that, in autonomic structures, IAA-RP may serve as a chemical mediator in complex circuits involved in blood pressure regulation and, more generally, sympathetic drive.

Two Cases of Spinocerebellar Ataxia Accompanied by Involvement of the Skeletal Motor Neuron System and Bulbar Palsy

We report two patients with spinocerebellar ataxia (SCA) with cranial and spinal motor neuron involvement. They initially presented with cerebellar ataxia, followed by bulbar palsy and limb motor neuron sign. One of the patients had a brother with allied disorder. SCA type 1 (SCA1), SCA3 and SCA6 have been reported to involve the motor neuron system, but they were excluded by DNA analyses in the present two patients. These two patients may form a distinct disease entity among SCAs.

Identification of genes necessary for the development of cranial motor neurons

Identification of genes necessary for the development of cranial motor neurons

A subunit of the mediator complex regulates vertebrate neuronal development

The unique profiles of gene expression dictate distinct cellular identity. How these profiles are established during development is not clear. Here we report that the mutant motionless (mot), identified in a genetic screen for mutations that affect neuronal development in zebrafish, displays deficits of monoaminergic neurons and cranial sensory ganglia, whereas expression of the pan-neuronal marker Hu is largely unperturbed; GABAergic and subsets of cranial motor neurons do not appear to be deficient. Positional cloning reveals that mot encodes Med12, a component of the evolutionarily conserved Mediator complex, whose in vivo function is not well understood in vertebrates. mot/med12 transcripts are enriched in the embryonic brain and appear distinct from two other Mediator components Med17 and Med21. Delivery of human med12 RNA into zebrafish restores normality to the mot mutant and, strikingly, leads to premature neuronal differentiation and an increased production of monoaminergic neuronal subtypes in WT. Further investigation reveals that mot/med12 is necessary to regulate, and when overexpressed is capable of increasing, the expression of distinct neuronal determination genes, including zash1a and lim1, and serves as an in vivo cofactor for Sox9 in this process. Together, our analyses reveal a regulatory role of Mot/Med12 in vertebrate neuronal development.

Longitudinal study of the motor response to levodopa in Parkinson's disease

In this prospective study of 34 patients with Parkinson's disease, measurements of the short duration levodopa motor response have been performed in defined off states at 3 yearly intervals over a mean period of 11.4 years from the point of commencement of levodopa treatment. Twenty-two patients were still available for study; 10 had died and 2 were lost to follow-up. The levodopa motor response amplitude increases over the first 5 years of treatment, and thereafter, on and off scores worsen in parallel with conservation of the response. Patients who developed motor fluctuations within the first 5 years of treatment had, on average, a stronger response to levodopa with significantly better on phase motor function (P = 0.003). Although the proportion of "midline" motor disability (affecting gait, balance, and cranial motor function) increases with time, these deficits do not actually become unresponsive to levodopa. Patients who developed dementia had a significantly more rapid decline in motor function. The latest graph of serial scores for the whole cohort shows an upward curving or exponential increase in motor disability after the first decade of treatment. Applying a notional untreated disability line to this graph--an estimate of the disability that would have accrued if drugs had never been given--we suggest that the long-duration response to levodopa eventually runs down with disease progression.

The axotomy‐induced neuropeptides galanin and pituitary adenylate cyclase‐activating peptide promote axonal sprouting of primary afferent and cranial motor neurones

The neuropeptides galanin and pituitary adenylate cyclase-activating peptide (PACAP) are markedly up-regulated in response to peripheral nerve lesion. Both peptides are involved in neuronal differentiation and neurite outgrowth during development. In this study, we investigated the effects of galanin and PACAP on axonal elongation and sprouting by adult rat sensory neurones in vitro and facial motor neurones in vivo. Dissociated rat dorsal root ganglion neurones were plated on laminin substrate and analysed morphometrically. Both the mean axonal length and the number of branch points significantly increased in the presence of galanin or PACAP (2-5 microm). Effects on axonal collateralization were investigated in the rat facial nerve lesion model by direct application of the peptides to collagen-filled conduits entubulating the transected facial nerve stumps. Triple retrograde labelling of brainstem neurones confirmed that the peptides potently induce axonal sprouting of cranial motor neurones. The number of neurones regenerating into identified rami of the facial nerve increased up to fivefold. Biometrical analysis of whisking behaviour revealed that galanin and PACAP impaired the functional outcome when compared with vehicle-treated animals 8 weeks after surgery. In conclusion, although galanin and PACAP have been established as neurotrophic molecules with respect to axonal development and regeneration, their potential as treatments for peripheral nerve lesions appears limited because of the extensive stimulation of collateral axon branching. These branches are misrouted towards incorrect muscles and cause impairment in their coordinated activity.

P3-174: The regional distribution and magnitude of the pathology in Alzheimer disease correlate with the regional pattern and intensity of the aminergic innervation of the forebrain

A fundamental, yet unexplained feature in Alzheimer disease (AD) is the selective vulnerability (“pathoklisis”). The allocortex including limbic nuclei, the neocortical association areas, the truncothalamic nuclei, the upper raphe nuclei and the locus ceruleus belong to the most severely affected brain regions. Conversely, the cerebellar cortex and its deep nuclei, the thalamic relay nuclei, many brainstem nuclei, as well as the cranial and spinal motor nuclei show little, if any, pathology. The aim of the present investigation was to examine semiquantitatively on the basis of data from the literature whether the magnitude of the ascending neuroaminergic innervation and receptor distribution might correlate with the regionally distinct distribution and severity of the pathology in the AD brain. Four aminergic systems with ascending projections, the cholinergic, serotoninergic, noradrenergic, and histaminergic systems, known to sustain severe pathology themselves in AD, were included in the evaluation. Neuroamine transmitter (NAT) levels and the expression of NAT receptors were evaluated for 235 CNS regions and nuclei semiquantitatively from data provided by more than 350 original papers, monographs and book articles. In order to standardize the very heterogeneously, i.e., both numerically or alpha–numerically, documented data values for pooling, a five category evaluation score was used. Categorization was accomplished by “numerical coding of semantics”: Semantically weighted descriptions were changed to numerical values according to the following evaluation score: (0) none, absent, almost none, very low, very few, single, scarce, scanty,etc.; (1) low, few, some, little, minor, scattered, infrequent, mild,etc.; (2) medium, intermediate, moderate, etc.; (3) high, marked, rich, strong, prominent, dense, intense,etc.; (4) abundant, large amounts, very strong, very high, highest, etc.. Intermediate values (.5) were permitted in case of descriptions like “medium to marked“ (=2.5). Numerical values displayed in tables were changed to four equal categories. Semiquantitative categorization of the pathology was accomplished accordingly. Indeed, a rough correlation can be shown between the magnitude of aminergic innervation (NAT levels and receptor densities), and the distribution and severity of the pathology: As a rule, severely affected (e.g., limbic) regions receive high while little affected (e.g.,cerebellar) regions receive low NAT innervation, with few exceptions.

Precraniate origin of cranial motoneurons.

The craniate head is innervated by cranial sensory and motor neurons. Cranial sensory neurons stem from the neurogenic placodes and neural crest and are seen as evolutionary innovations crucial in fulfilling the feeding and respiratory needs of the craniate "new head." In contrast, cranial motoneurons that are located in the hindbrain and motorize the head have an unclear phylogenetic status. Here we show that these motoneurons are in fact homologous to the motoneurons of the sessile postmetamorphic form of ascidians. The motoneurons of adult Ciona intestinalis, located in the cerebral ganglion and innervating muscles associated with the huge "branchial basket," express the transcription factors CiPhox2 and CiTbx20, whose vertebrate orthologues collectively define cranial motoneurons of the branchiovisceral class. Moreover, Ciona's postmetamorphic motoneurons arise from a hindbrain set aside during larval life and defined as such by its position (caudal to the prosensephalic sensory vesicle) and coexpression of CiPhox2 and CiHox1, whose orthologues collectively mark the vertebrate hindbrain. These data unveil that the postmetamorphic ascidian brain, assumed to be a derived feature, in fact corresponds to the vertebrate hindbrain and push back the evolutionary origin of cranial nerves to before the origin of craniates.

First a third, then a fourth nerve palsy in multiple intracranial aneurysms

Intracranial aneurysms may be a cause of ocular motor dysfunction, the third nerve being more often involved than the two other cranial ocular motor nerves. We report the unusual occurrence of an isolated fourth nerve palsy related to a cavernous carotid aneurysm, angiographically confirmed. The patient already underwent clipping of a ruptured posterior communicating artery aneurysm 17 years earlier, revealed at that time by a third nerve palsy. Endovascular treatment of the cavernous carotid aneurysm was performed, resulting in complete recovery of the palsy. Occurrence of an acquired fourth nerve palsy in a patient with a past history of aneurysm should prompt neurovascular imaging as multiple aneurysms may cause sequential ocular motor palsies.

Localization and ontogeny of damage-induced neuronal endopeptidase mRNA-expressing neurons in the rat nervous system

Neuropeptides are crucial mediators in nervous and endocrine systems. Processing and degradation, the major regulatory mechanisms, of enzymes are essential for the control of these peptidergic intercellular signaling systems. Damage-induced neuronal endopeptidase (or endothelin converting enzyme-like1), a member of the neprilysin family, has recently been identified as an M13 zinc metalloprotease. Damage-induced neuronal endopeptidase mRNA expression is strikingly restricted to neurons, and is remarkably induced in response to various types of neuronal injuries, although its function and substrate remain unknown. To clarify the role of damage-induced neuronal endopeptidase, we examined the localization and ontogeny of damage-induced neuronal endopeptidase mRNA expression in the rat nervous system using in situ hybridization. Damage-induced neuronal endopeptidase mRNA was detected at embryonic day 12, and its expression restricted to the ventral region of the neural tube. Subsequently, expression was also apparent in primordia of the striatum, hypothalamus, and cranial motor nuclei during neural development. This specific distribution was relatively maintained in the adult brain, although expression levels became weaker. Expression of damage-induced neuronal endopeptidase was absent in the cerebral cortex, hippocampus, and cerebellum. In addition to prominent expression in CNS, intestinal and sensory ganglia and retina demonstrated transient intense damage-induced neuronal endopeptidase mRNA expression during the embryonic period that then declined, and disappeared after birth. The results indicated that damage-induced neuronal endopeptidase might play an important role in embryonic neural development, in particular in peripheral ganglia derived from the neural crest, and in some neurons originating from the basal plate such as the hypothalamus and cranial motor neurons.

The importance of brainstem mapping in brainstem surgical anatomy before the fourth ventricle and implication for intraoperative neurophysiological mapping

Brain stem mapping (BSM) is an intraoperative neurophysiological procedure to localize cranial motor nuclei on the floor of the fourth ventricle. BSM enables neurosurgeon to understand functional anatomy on the distorted floor of the fourth ventricle, thus, it is emerging as an indispensable tool for challenging brain stem surgery. The authors described the detail of BSM with the special emphasis on its clinical application for the brain stem lesion. Surgical implications based on the result of brains stem mapping would be also informative before planning a brain stem surgery through the floor of fourth ventricle. Despite the recent advancement of MRI to depict the lesion in the brain stem, BSM remains as the only way to provide surgical anatomy in the operative field. BSM could guide a neurosurgeon to the inside of brain stem while preventing direct damage to the cranial motor nuclei on the floor of the fourth ventricle. It is expected that understanding its advantage and limitations would help neurosurgeon to perform safer surgery to the brain stem lesion.

A Genetic Screen for Mutations That Affect Cranial Nerve Development in the Mouse

Cranial motor and sensory nerves arise stereotypically in the embryonic hindbrain, act as sensitive indicators of general and region-specific neuronal development, and are directly or indirectly affected in many human disorders, particularly craniofacial syndromes. The molecular genetic hierarchies that regulate cranial nerve development are mostly unknown. Here, we describe the first mouse genetic screen that has used direct immunohistochemical visualization methods to systematically identify genetic loci required for cranial nerve development. After screening 40 pedigrees, we recovered seven new neurodevelopmental mutations. Two mutations model human genetic syndromes. Mutation 7-1 causes facial nerve anomalies and a reduced lower jaw, and is located in a region of conserved synteny with an interval associated with the micrognathia and mental retardation of human cri-du-chat syndrome. Mutation 22-1 is in the Pax3 gene and, thus, models human Waardenburg syndrome. Three mutations cause global axon guidance deficits: one interferes with initial motor axon extension from the neural tube, another causes overall axon defasciculation, and the third affects general choice point selection. Another two mutations affect the oculomotor nerve specifically. Oculomotor nerve development, which is disrupted by six mutations, appears particularly sensitive to genetic perturbations. Phenotypic comparisons of these mutants identifies a "transition zone" that oculomotor axons enter after initial outgrowth and in which new factors govern additional progress. The number of interesting neurodevelopmental mutants revealed by this small-scale screen underscores the promise of similar focused genetic screens to contribute significantly to our understanding of cranial nerve development and human craniofacial syndromes.

Projections of the ventrolateral pontine vocalization area in the squirrel monkey

In four squirrel monkeys (Saimiri sciureus), the tracer biotin dextranamine (BDA) was injected into the ventrolateral pons at a site at which injection of the glutamate antagonist kynurenic acid blocked vocalization electrically elicited from the periaqueductal gray (PAG). Anterograde projections could be traced into all cranial motor and sensory nuclei involved in phonation, that is, the nucleus ambiguus, facial, hypoglossal and trigeminal motor nuclei, the motorneuron column in the ventral gray substance innervating the extrinsic laryngeal muscles, the nucleus retroambiguus, solitary tract and spinal trigeminal nuclei. Projections were also found into a number of auditory nuclei, namely the nucleus cochlearis-complex, superior olive, ventral and dorsal nuclei of the lateral lemniscus and inferior colliculus. Furthermore, there were projections into the reticular formation of the lateral and dorsocaudal medulla and lateral pons, into nucleus gracilis, inferior and medial vestibular nuclei, lateral reticular nucleus, ventral raphe, pontine gray, superior colliculus, PAG and mediodorsal thalamic nucleus. Injection of the tracer wheat germ agglutinin-conjugated horseradish peroxidase into the ventrolateral pontine vocalization-blocking area in one animal yielded retrograde labeling throughout the PAG. Injection of BDA into a vocalization-eliciting site of the PAG in another animal yielded projections into the ventrolateral pontine vocalization-blocking area. It is concluded that the ventral paralemniscal area in the ventrolateral pons represents a relay station of the descending periaqueductal vocalization-controlling pathway.

Slit-mediated repulsion is a key regulator of motor axon pathfinding in the hindbrain

The floor plate is known to be a source of repellent signals for cranial motor axons, preventing them from crossing the midline of the hindbrain. However, it is unknown which molecules mediate this effect in vivo. We show that Slit and Robo proteins are candidate motor axon guidance molecules, as Robo proteins are expressed by cranial motoneurons, and Slit proteins are expressed by the tissues that delimit motor axon trajectories, i.e. the floor plate and the rhombic lip. We present in vitro evidence showing that Slit1 and Slit2 proteins are selective inhibitors and repellents for dorsally projecting, but not for ventrally projecting, cranial motor axons. Analysis of mice deficient in Slit and Robo function shows that cranial motor axons aberrantly enter the midline, while ectopic expression of Slit1 in chick embryos leads to specific motor axon projection errors. Expression of dominant-negative Robo receptors within cranial motoneurons in chick embryos strikingly perturbs their projections, causing some motor axons to enter the midline, and preventing dorsally projecting motor axons from exiting the hindbrain. These data suggest that Slit proteins play a key role in guiding dorsally projecting cranial motoneurons and in facilitating their neural tube exit.

Gli function is essential for motor neuron induction in zebrafish

The Gli family of zinc-finger transcription factors mediates Hedgehog (Hh) signaling in all vertebrates. However, their roles in ventral neural tube patterning, in particular motor neuron induction, appear to have diverged across species. For instance, cranial motor neurons are essentially lost in zebrafish detour ( gli1 − ) mutants, whereas motor neuron development is unaffected in mouse single gli and some double gli knockouts. Interestingly, the expression of some Hh-regulated genes ( ptc1, net1a, gli1) is mostly unaffected in the detour mutant hindbrain, suggesting that other Gli transcriptional activators may be involved. To better define the roles of the zebrafish gli genes in motor neuron induction and in Hh-regulated gene expression, we examined these processes in you-too ( yot) mutants, which encode dominant repressor forms of Gli2 (Gli2 DR), and following morpholino-mediated knockdown of gli1, gli2, and gli3 function. Motor neuron induction at all axial levels was reduced in yot ( gli2 DR ) mutant embryos. In addition, Hh target gene expression at all axial levels except in rhombomere 4 was also reduced, suggesting an interference with the function of other Glis. Indeed, morpholino-mediated knockdown of Gli2 DR protein in yot mutants led to a suppression of the defective motor neuron phenotype. However, gli2 knockdown in wild-type embryos generated no discernable motor neuron phenotype, while gli3 knockdown reduced motor neuron induction in the hindbrain and spinal cord. Significantly, gli2 or gli3 knockdown in detour ( gli1 − ) mutants revealed roles for Gli2 and Gli3 activator functions in ptc1 expression and spinal motor neuron induction. Similarly, gli1 or gli3 knockdown in yot ( gli2 DR ) mutants resulted in severe or complete loss of motor neurons, and of ptc1 and net1a expression, in the hindbrain and spinal cord. In addition, gli1 expression was greatly reduced in yot mutants following gli3, but not gli1, knockdown, suggesting that Gli3 activator function is specifically required for gli1 expression. These observations demonstrate that Gli activator function (encoded by gli1, gli2, and gli3) is essential for motor neuron induction and Hh-regulated gene expression in zebrafish.

Dual roles of zygotic and maternal Scribble1 in neural migration and convergent extension movements in zebrafish embryos

In the developing vertebrate hindbrain, the characteristic trajectory of the facial (nVII) motor nerve is generated by caudal migration of the nVII motor neurons. The nVII motor neurons originate in rhombomere (r) 4, and migrate caudally into r6 to form the facial motor nucleus. In this study, using a transgenic zebrafish line that expresses green fluorescent protein (GFP) in the cranial motor neurons, we isolated two novel mutants, designated landlocked (llk) and off-road (ord), which both show highly specific defects in the caudal migration of the nVII motor neurons. We show that the landlocked locus contains the gene scribble1 (scrb1), and that its zygotic expression is required for migration of the nVII motor neurons mainly in a non cell-autonomous manner. Taking advantage of the viability of the llk mutant embryos, we found that maternal expression of scrb1 is required for convergent extension (CE) movements during gastrulation. Furthermore, we show a genetic interaction between scrb1 and trilobite(tri)/strabismus(stbm) in CE. The dual roles of the scrb1 gene in both neuronal migration and CE provide a novel insight into the underlying mechanisms of cell movement in vertebrate development.